Overview

  • Product name
    Anti-TTF1 antibody [8G7G3/1]
    See all TTF1 primary antibodies
  • Description
    Mouse monoclonal [8G7G3/1] to TTF1
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, IHC-Fr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Dog
  • Immunogen

    Recombinant full length protein corresponding to Rat TTF1.
    Database link: P43699

  • Positive control
    • IHC-P: Human lung, thyroid and lung adenocarcinoma tissue. IHC-Fr: Mouse thyroid tissue. Flow cytometry: A549 cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity
    Protein A/G purified
  • Purification notes
    Purified from TCS by protein A/G.
  • Clonality
    Monoclonal
  • Clone number
    8G7G3/1
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab72876 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/50. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

PMID 20081193

IHC-Fr 1/50.
Flow Cyt Use 20µl for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Transcription factor that binds and activates the promoter of thyroid specific genes such as thyroglobulin, thyroperoxidase, and thyrotropin receptor. Crucial in the maintenance of the thyroid differentiation phenotype. May play a role in lung development and surfactant homeostasis.
  • Tissue specificity
    Thyroid and lung.
  • Involvement in disease
    Defects in NKX2-1 are the cause of benign hereditary chorea (BHC) [MIM:118700]; also known as hereditary chorea without dementia. BHC is an autosomal dominant movement disorder. The early onset of symptoms (usully before the age of 5) and the observation that in some BHC families the symptoms tend to decrease in adulthood suggests that the disorder results from a developmental disturbance of the brain. BHC is non-progressive and patients have normal or slightly below normal intelligence. There is considerable inter- and intrafamilial variability, including dysarthria, axial distonia and gait disturbances.
    Defects in NKX2-1 are the cause of choreoathetosis, hypothyroidism, and neonatal respiratory distress (CHNRD) [MIM:610978]. This syndrome include neurological, thyroid, and respiratory problems.
  • Sequence similarities
    Belongs to the NK-2 homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Post-translational
    modifications
    Phosphorylated on serine residues.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • AV026640 antibody
    • BCH antibody
    • Benign chorea antibody
    • BHC antibody
    • Homeobox protein NK 2 homolog A antibody
    • Homeobox protein NK-2 homolog A antibody
    • Homeobox protein Nkx 2.1 antibody
    • Homeobox protein Nkx-2.1 antibody
    • Homeobox protein Nkx2.1 antibody
    • NK 2 antibody
    • NK 2 homolog A antibody
    • NK2 antibody
    • NK2 homeobox 1 antibody
    • NK2, drosophila, homolog of, A antibody
    • NK2.1, mouse, homolog of antibody
    • Nkx 2 1 antibody
    • NKX 2.1 antibody
    • NKX 2A antibody
    • NKX2 1 antibody
    • Nkx2-1 antibody
    • NKX2.1 antibody
    • NKX21_HUMAN antibody
    • NKX2A antibody
    • T EBP antibody
    • T/EBP antibody
    • TEBP antibody
    • Thyroid nuclear factor 1 antibody
    • Thyroid nuclear factor antibody
    • Thyroid specific enhancer binding protein antibody
    • Thyroid transcription factor 1 antibody
    • Tin man antibody
    • Tinman antibody
    • TITF 1 antibody
    • TITF1 antibody
    • TTF 1 antibody
    • TTF-1 antibody
    • TTF1 antibody
    see all

Images

  • Formalin-fixed, paraffin-embedded human lung tissue stained for TTF1 with ab72876 at 1/50 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human lung adenocarcinoma tissue stained for TTF1 with ab72876 at 1/50 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human thyroid tissue stained for TTF1 with ab72876 at 1/50 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human lung tissue stained for TTF1 with ab72876 at 1/50 dilution in immunohistochemical analysis.

  • ab72876 staining TTF1 in Mouse thyroid tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde, permeabilized with 0.1% Tween 20 and blocked with 10% serum for 30 minutes at 24°C. Samples were incubated with primary antibody (1/50 in 10% goat serum in PBS) for 12 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • ab72876 at 1/50 dilution staining TTF1 in human lung by Immunohistochemistry, Formalin-fixed, Paraffin-embedded tissue.
  • Overlay histogram showing A549 cells stained with ab72876 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab72876, 20µl/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A549 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Kortlever RM  et al. Myc Cooperates with Ras by Programming Inflammation and Immune Suppression. Cell 171:1301-1315.e14 (2017). Read more (PubMed: 29195074) »
  • Zhu L  et al. Surgical monotherapy may be a suitable therapeutic strategy for advanced collecting (Bellini) duct carcinoma: A case report and literature review. Exp Ther Med 12:1181-1184 (2016). IHC-P ; Human . Read more (PubMed: 27446340) »
See all 5 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (A549)
Permeabilization
Yes - 0.1% Tween 20 and 0.1% Triton X-100 in PBS + 2% FBS
Specification
A549
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: 4°C
Fixative
Paraformaldehyde

Heather Wanczyk

Verified customer

Submitted Jun 05 2018

Question
Answer

The concentration for ab72876 Lot# GR143259-6 is 0.52mg/ml.

Read More

Question
Answer

The concentration of ab72876, Anti-TTF1 antibody [8G7G3/1] , lot#GR105660-1 is 0.52 mg/ml.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Mouse Tissue sections (mouse thyroid)
Specification
mouse thyroid
Permeabilization
Yes - 0.1% Tween 20
Fixative
Paraformaldehyde

Mr. Sanjay Gawade

Verified customer

Submitted Jun 26 2013

Question

Kindly find the answers to the questionnaire.

Questionnaire:

General Information
Antibody storage conditions (temperature/reconstitution etc) - 4 C in dark.


Description of the problem (high background, low signal, no signal etc.)- no signal at all.


Sample (Species/Cell type/Cell line etc.)- Thyroid tissue from mice.


Sample preparation for single cell preparation (Buffer etc.)- thyroid tissue from mice was digested either by Collagenase D or Liberase. Washing was carried using either PBS 2% FCS or IMDM 10% FCS when cells were alive and when fixed were washed by wash buffer from ebioscience.


Number of cells used -100000.


Permeabilization, fixation step - Fixation by either 4% paraformaldehyde frehly made 10 mins at room temperature or by using fixation buffer from ebioscience which also has 4% formaldehyde or using 80% methanol for 5 mins on ice. Permeabilization was carried out using permeabilization buffer from ebioscience.

Blocking conditions (Buffer/time period, Blocking agent etc.)-
10% normal goat serum in 1X PBS containing 0.3M glycine .

Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) - ab72876 incubated at room temperature or on ice after fixation and permeabilization for 30 mins or 45 mins.

Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)- goat antirabbit from biolegend at1:500 dilution with A647 conjugation for 30 mins at room temperature.

Detection method- FACS Aria from BD.

Positive and negative controls used (please specify) - positive control - thyroid tissue from mice. Negative control - adult thymus or spleen from mice.

Optimization attempts (problem solving)
How many times have you tried the FACS? 4 times.


Have you run a "No Primary" control? yes


Yes No
Do you obtain the same results every time? yes
Yes No

What steps have you altered? changed fixation agent, used 4% paraformaldehyde instead of 80% methanol. changed primary and secondary antibody incubation times. I also used goat antirabbit secondary antibody with A 488 conjugation.


Additional Notes:


Data:
We would appreciate if you are able to provide any data/histograms which will help us to assess the results

Read More
Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that ab72876 and ab53490 are tested and covered by our 6 month guarantee for use in FlowCytometry and mouse samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would like to offer some suggestions to help optimise the results from this antibodies. I would also appreciate if you can confirm some further details:

1. Could you please confirm the order number and date of purchase for the two antibodies? This will make sure that I can offer some compensation in case the antibodies do not work as cited on the datasheet.

2. We would recommend it would be beneficial to check the quality of the sample by staining with trypan blue to confirm the viability of samples is over 95%.

3. Could you confirm the concentration of antibody that was used? We recommend to use 20µl for 106 cells with the ab72876.

4. Could you confirm that the goat anti rabbit antibodies were used for both experiments. The primary antibodies ab72876 and ab53490 are mouse monoclonal antibodies. For this reason we would regrettably not expect them to be detected by an anti rabbit antibody. We would recommend to use a secondary antibody raised against mouse in this case.

I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
I would like to reassure you that this antibody is tested and covered by our 6 month guarantee for FlowCytometry and mouse, rat and human samples. In the event that a product is not functioning in the tested applications and species cited on the product data sheet (and the problem has been reported within 6 months of purchase), we will be pleased to provide a credit note, free of charge replacement or refund.
I would like to investigate this particular case further for you, and also obtain more information for our quality monitoring records. In order to proceed with this, I have enclosed a technical questionnaire below. I would appreciate if you could complete this. It will help you put the information we require together very easily.
I would appreciate if you could also provide an image which would help us to assess the results.
Thank you for your time and cooperation. We look forward to receiving the completed questionnaire.



Additionally I like to send you the protocol that was used for the FlowCytometry testing as cited on the datasheet of ab72876 beside the provided image for FlowCytometry.



A549 cells stained with ab72876 (red line).
Cells were fixed with 4% paraformaldehyde (10 min)
then permeabilized with 0.1% PBS-Tween for 20 min
incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions
followed by the antibody (ab72876, 20µl/1x106 cells) for 30 min at 22ºC
The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (https://www.abcam.com/ab96879.html) at 1/500 dilution for 30 min at 22ºC
Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (https://www.abcam.com/ab91353.html, 2µg/1x106 cells) used under the same conditions
Acquisition of >5,000 events was performed
This antibody gave a positive signal in A549 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.





Questionnaire:

Order Details
Antibody code:

Problem:
Choose: Non-specific staining No signal or weak signal High background

Lot number

Purchase order number
or preferably Abcam order number

General Information
Antibody storage conditions (temperature/reconstitution etc)


Description of the problem (high background, low signal, no signal etc.)


Sample (Species/Cell type/Cell line etc.)


Sample preparation for single cell preparation (Buffer etc.)


Number of cells used


Permeabilization, fixation step


Blocking conditions (Buffer/time period, Blocking agent etc.)


Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step)


Detection method


Positive and negative controls used (please specify)



Optimization attempts (problem solving)
How many times have you tried the FACS?


Have you run a "No Primary" control?

Yes No
Do you obtain the same results every time?
Yes No

What steps have you altered?


Additional Notes:

Data:
We would appreciate if you are able to provide any data/histograms which will help us to assess the results

Read More

Question
Answer

Thank you for contacting us.

The concentration of lot GR76703-1 is 0.43 mg/ml.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Answer

Merci pour ce complément d'informations qui va nous permettre de chercher l'origine du problème rencontré avec le ab40880.

Comme convenu j'ai mis en place l'envoi d'une unité de ab72876, le numéro de commande de remplacement gratuit de ce produit est *****.
Par mesure préventive, j'ai pris l'initiative de retarder l'envoi du ab72876 à lundi afin qu'il ne reste pas en transit tout le weekend. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

Veuillez noter que cet anticorps de remplacement est également couvert par la garantie Abpromise, donc n'hésitez pas à nous contacter de nouveau si nécessaire.

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Answer

Thank you for contacting us.

The concentration of the lot you have is 0.43 mg/ml.

We have no western blotting data and the application was added to the datasheet based on a mistaken assumption that the following reference used the antibody in this application.

Manuel M et al. The transcription factor Foxg1 regulates the competence of telencephalic cells to adopt subpallial fates in mice. Development 137:487-97 (2010).

It appears to me that the antibody was only used for IHC, though, so we will remove western blotting from the datasheet.

If you do decide to use the antibody for western blotting, we will still honor our guarantee. I recommend trying a 1/200 dilution, then adjusting for subsequent blots if necessary. If the results are not satisfactory, please contact us.

The antibody has not, to our knowledge, been tested in any kind of ELISA. For an indirect ELISA, I recommend testing a range of dilutions, for instance 1/100, 1/500, and 1/2500.

Read More

Answer

Thank you for contacting us. Based on the Uniprot information for Rat TTF1 (http://www.uniprot.org/uniprot/P23441), the expected weight based on sequence is around 35 kDa. Real world weight may differ due to factors such as post translational modification, relative charge or other factors. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews

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